US4816444AExpiredUtility

Cell growth inhibitory substance

98
Assignee: UNIV ARIZONA STATEPriority: Jul 10, 1987Filed: Jul 10, 1987Granted: Mar 28, 1989
Est. expiryJul 10, 2007(expired)· nominal 20-yr term from priority
A61P 35/00A61K 38/00C07K 5/0205
98
PatentIndex Score
297
Cited by
2
References
18
Claims

Abstract

A potent cell growth inhibitory substance obtained from the Indian Ocean sea hare Dolabella, herein denominated "dolastatin 10"; and pharmaceutical preparations containing same; and methods useful for the treatment of a host afflicted with neoplastic disease therewith.

Claims

exact text as granted — not AI-modified
Accordingly, what is claimed is: 
     
       1. A cell growth inhibitory substance denominated dolastatin 10 and having the structural formula: ##STR5## 
     
     
       2. A pharmaceutical preparation comprising a pharmaceutically acceptable carrier and an effective amount of a natural or synthetic cell growth inhibitory substance or a non-toxic pharmaceutical active derivative thereof, said substance having the structural formula: ##STR6## 
     
     
       3. A cell growth inhibitory substance according to claim 1 having the following N.M.R. characteristics:   ______________________________________                                    
Chemical Shift, ppm                                                       
Structure          .sup.1 H(mult; J,Hz;                                   
Assignment                                                                
        13C(mult)  Integration)     NOE                                   
______________________________________                                    
 2      170.51(s)  7.717(d; 3.3; 1H)                                      
 4      142.77(d)  7.254(d; 3.3; 1H)                                      
 5      118.76(d)  5.516(ddd; 5.7,7.2, 9.3; 1H)                           
 6       53.02(d)  3.399(dd; 5.7,14; 1H)                                  
 6a      41.48(t)  3.256(dd; 5.7; 14; 1H)                                 
 6b1    137.74(s)  7.243(d; 7.9; 2H)                                      
 6b2,6b6                                                                  
        128.74(d)×2                                                 
                   7.214(dd; 7.9,9.2; 2H)                                 
 6b3,6b5                                                                  
        129.80(d)×2                                                 
                   7.194(t; 9.2; 1H)                                      
 6b4    127.02(d)  7.256(t; 7.6; 1H)                                      
 7                                  9,9a,10,                              
                                    10ab,11                               
 8      175.67(s)  2.282(quintet; 7.2; 1H)                                
                                    7                                     
 9       44.79(d)  1.085(d; 7.1; 3H)                                      
 9a      14.49(q)  3.845(dd; 2.0,8.2; 1H)                                 
                                    7                                     
10       82.05(d)  3.309(s; 3H)     6,7                                   
10ab     60.89(q)  3.985(m; 1H)                                           
11       59.86(d)  1.804(ddd; 5.5,7.0; 19; 1H)                            
12       25.00(t)  1.608(ddd; 7; 9.2,19; 1H)                              
                   1.446(ddd; 4.7,7,19; 1H)                               
13       25.45(t)  1.715(ddd; 4.7,7.8,12.7; 1H)                           
                   3.401(dd; 7.8,10; 1H)                                  
                                    17                                    
14       48.03(t)  3.390(m; 1H).sup.b                                     
16      174.01(s)                                                         
17       38.11(t)  2.394(ABq; 9.0; 2H)                                    
                                    14                                    
18       78.86(d)  4.122(broad t; 8.7; 1H)                                
18ab     58.16(q)  3.313(s; 3H)     22                                    
19       54.11(d)  3.26-3.39(1H).sup.c                                    
19a      33.62(d)  1.680(1H).sup.b                                        
19b      26.25(t)  1.370(broad m; 1H)                                     
                   1.000(broad m; 1H).sup.b                               
19c      10.92(q)  0.823(t; 7.4; 3H)                                      
19d      19.82(q)  1.003(d; 6.8; 3H)                                      
20a      30.09(q)  3.012(s; 3H)     22                                    
21      171.39(s)                                                         
22       54.20(d)  4.761(dd; 6.5, 8.8; 1H)                                
                                    18,18ab,                              
                                    20,19                                 
22a      31.42(d)  1.983(sextet; 6.7; 1H)                                 
22b      18.18(q)  0.941(d; 6.8; 3H)                                      
22c      16.09(q)  0.977(d; 6.8; 3H)                                      
23                 6.861(d; 8.9; 1H)                                      
                                    25bc                                  
24      172.44(s)                                                         
25       76.77(d)  2.454(d; 6.9; 1H)                                      
                                    23                                    
25bc     49.92(q)×2                                                 
                   2.262(s; 6H)     23                                    
26       28.08(d)  2.073(sextet; 6.7; 1H)                                 
27       20.24(q)  0.964(d; 6.8; 3H)                                      
28       18.18(q)  0.902(d; 6.8; 3H)                                      
______________________________________                                    
 .sup.a Residual CHDCl.sub.2 as internal reference (5.32 ppm).            
 .sup.b Overlapping signal.                                               
 .sup.c Signal assigned from NOE data.                                    
     
     
     
       4. A pharmaceutical preparation according to claim 2 in which said substance has the following N.M.R. characteristics:   ______________________________________                                    
Chemical Shift, ppm                                                       
Structure          .sup.1 H(mult; J,Hz;                                   
Assignment                                                                
        13C(mult)  Integration)     NOE                                   
______________________________________                                    
 2      170.51(s)  7.717(d; 3.3; 1H)                                      
 4      142.77(d)  7.254(d; 3.3; 1H)                                      
 5      118.76(d)  5.516(ddd; 5.7,7.2, 9.3; 1H)                           
 6       53.02(d)  3.399(dd; 5.7,14; 1H)                                  
 6a      41.48(t)  3.256(dd; 5.7; 14; 1H)                                 
 6b1    137.74(s)  7.243(d; 7.9; 2H)                                      
 6b2,6b6                                                                  
        128.74(d)×2                                                 
                   7.214(dd; 7.9,9.2; 2H)                                 
 6b3,6b5                                                                  
        129.80(d)×2                                                 
                   7.194(t; 9.2; 1H)                                      
 6b4    127.02(d)  7.256(t; 7.6; 1H)                                      
 7                                  9,9a,10,                              
                                    10ab,11                               
 8      175.67(s)  2.282(quintet; 7.2; 1H)                                
                                    7                                     
 9       44.79(d)  1.085(d; 7.1; 3H)                                      
 9a      14.49(q)  3.845(dd; 2.0,8.2; 1H)                                 
                                    7                                     
10       82.05(d)  3.309(s; 3H)     6,7                                   
10ab     60.89(q)  3.985(m; 1H)                                           
11       59.86(d)  1.804(ddd; 5.5,7.0; 19; 1H)                            
12       25.00(t)  1.608(ddd; 7; 9.2,19; 1H)                              
                   1.446(ddd; 4.7,7,19; 1H)                               
13       25.45(t)  1.715(ddd; 4.7,7.8,12.7; 1H)                           
                   3.401(dd; 7.8,10; 1H)                                  
                                    17                                    
14       48.03(t)  3.390(m; 1H).sup.b                                     
16      174.01(s)                                                         
17       38.11(t)  2.394(ABq; 9.0; 2H)                                    
                                    14                                    
18       78.86(d)  4.122(broad t; 8.7; 1H)                                
18ab     58.16(q)  3.313(s; 3H)     22                                    
19       54.11(d)  3.26-3.39(1H).sup.c                                    
19a      33.62(d)  1.680(1H).sup.b                                        
19b      26.25(t)  1.370(broad m; 1H)                                     
                   1.000(broad m; 1H).sup.b                               
19c      10.92(q)  0.823(t; 7.4; 3H)                                      
19d      19.82(q)  1.003(d; 6.8; 3H)                                      
20a      30.09(q)  3.012(s; 3H)     22                                    
21      171.39(s)                                                         
22       54.20(d)  4.761(dd; 6.5, 8.8; 1H)                                
                                    18,18ab,                              
                                    20,19                                 
22a      31.42(d)  1.983(sextet; 6.7; 1H)                                 
22b      18.18(q)  0.941(d; 6.8; 3H)                                      
22c      16.09(q)  0.977(d; 6.8; 3H)                                      
23                 6.861(d; 8.9; 1H)                                      
                                    25bc                                  
24      172.44(s)                                                         
25       76.77(d)  2.454(d; 6.9; 1H)                                      
                                    23                                    
25bc     49.92(q)×2                                                 
                   2.262(s; 6H)     23                                    
26       28.08(d)  2.073(sextet; 6.7; 1H)                                 
27       20.24(q)  0.964(d; 6.8; 3H)                                      
28       18.18(q)  0.902(d; 6.8; 3H)                                      
______________________________________                                    
 .sup.a Residual CHDCl.sub.2 as internal reference (5.32 ppm).            
 .sup.b Overlapping signal.                                               
 .sup.c Signal assigned from NOE data.                                    
     
     
     
       5. A method of treating a host afflicted with neoplastic disease comprising administering to said host an effective amount of a natural or synthetic cell growth inhibitory substance or a pharmaceutically active non-toxic derivative thereof, said substance having the structural formula: ##STR7## 
     
     
       6. A method according to claim 5 in which said substance is administered intravenously, at a dosage level of from 0.1 up to about 20 mg per kilogram of host body weight. 
     
     
       7. A method according to claim 5 in which said substance is administered subcutaneously, at a dosage level of from 1 up to about 50 mg per kilogram of host body weight. 
     
     
       8. A method according to claim 5 in which said substance is administered orally, at a dosage level of from 5 up to about 100 mg per kilogram of host body weight. 
     
     
       9. A method according to claim 5 in which said neoplastic disease is lymphocytic leukemia P388. 
     
     
       10. A method according to claim 9 in which said small but effective amount comprises from about 1 up to about 4 μg per kilogram of host body weight. 
     
     
       11. A method according to claim 5 in which said substance has the following N.M.R. characteristics:   ______________________________________                                    
Chemical Shift, ppm                                                       
Structure          .sup.1 H(mult; J,Hz;                                   
Assignment                                                                
        13C(mult)  Integration)     NOE                                   
______________________________________                                    
 2      170.51(s)  7.717(d; 3.3; 1H)                                      
 4      142.77(d)  7.254(d; 3.3; 1H)                                      
 5      118.76(d)  5.516(ddd; 5.7,7.2, 9.3; 1H)                           
 6       53.02(d)  3.399(dd; 5.7,14; 1H)                                  
 6a      41.48(t)  3.256(dd; 5.7; 14; 1H)                                 
 6b1    137.74(s)  7.243(d; 7.9; 2H)                                      
 6b2,6b6                                                                  
        128.74(d)×2                                                 
                   7.214(dd; 7.9,9.2; 2H)                                 
 6b3,6b5                                                                  
        129.80(d)×2                                                 
                   7.194(t; 9.2; 1H)                                      
 6b4    127.02(d)  7.256(t; 7.6; 1H)                                      
 7                                  9,9a,10,                              
                                    10ab,11                               
 8      175.67(s)  2.282(quintet; 7.2; 1H)                                
                                    7                                     
 9       44.79(d)  1.085(d; 7.1; 3H)                                      
 9a      14.49(q)  3.845(dd; 2.0,8.2; 1H)                                 
                                    7                                     
10       82.05(d)  3.309(s; 3H)     6,7                                   
10ab     60.89(q)  3.985(m; 1H)                                           
11       59.86(d)  1.804(ddd; 5.5,7.0; 19; 1H)                            
12       25.00(t)  1.608(ddd; 7; 9.2,19; 1H)                              
                   1.446(ddd; 4.7,7,19; 1H)                               
13       25.45(t)  1.715(ddd; 4.7,7.8,12.7; 1H)                           
                   3.401(dd; 7.8,10; 1H)                                  
                                    17                                    
14       48.03(t)  3.390(m; 1H).sup.b                                     
16      174.01(s)                                                         
17       38.11(t)  2.394(ABq; 9.0; 2H)                                    
                                    14                                    
18       78.86(d)  4.122(broad t; 8.7; 1H)                                
18ab     58.16(q)  3.313(s; 3H)     22                                    
19       54.11(d)  3.26-3.39(1H).sup.c                                    
19a      33.62(d)  1.680(1H).sup.b                                        
19b      26.25(t)  1.370(broad m; 1H)                                     
                   1.000(broad m; 1H).sup.b                               
19c      10.92(q)  0.823(t; 7.4; 3H)                                      
19d      19.82(q)  1.003(d; 6.8; 3H)                                      
20a      30.09(q)  3.012(s; 3H)     22                                    
21      171.39(s)                                                         
22       54.20(d)  4.761(dd; 6.5, 8.8; 1H)                                
                                    18,18ab,                              
                                    20,19                                 
22a      31.42(d)  1.983(sextet; 6.7; 1H)                                 
22b      18.18(q)  0.941(d; 6.8; 3H)                                      
22c      16.09(q)  0.977(d; 6.8; 3H)                                      
23                 6.861(d; 8.9; 1H)                                      
                                    25bc                                  
24      172.44(s)                                                         
25       76.77(d)  2.454(d; 6.9; 1H)                                      
                                    23                                    
25bc     49.92(q)×2                                                 
                   2.262(s; 6H)     23                                    
26       28.08(d)  2.073(sextet; 6.7; 1H)                                 
27       20.24(q)  0.964(d; 6.8; 3H)                                      
28       18.18(q)  0.902(d; 6.8; 3H)                                      
______________________________________                                    
 .sup.a Residual CHDCl.sub.2 as internal reference (5.32 ppm).            
 .sup.b Overlapping signal.                                               
 .sup.c Signal assigned from NOE data.                                    
     
     
     
       12. A method according to claim 5 in which said neoplastic disease is murine melanoma. 
     
     
       13. A method according to claim 12 in which said small but effective amount comprises from about 1.44 up to about 11.1 μg per kilogram of host body weight. 
     
     
       14. A method according to claim 5 in which said neoplastic disease is human melanoma. 
     
     
       15. A method according to claim 14 in which said small but effective amount comprises from about 3.25 up to about 26 μg per kilogram of host body weight. 
     
     
       16. A method according to claim 11 in which said substance is administered intravenously, at a dosage level of from 0.1 up to about 20 mg per kilogram of host body weight. 
     
     
       17. A method according to claim 11 in which said substance is administered subcutaneously, at a dosage level of from 1 up to about 50 mg per kilogram of host body weight. 
     
     
       18. A method according to claim 11 in which said substance is administered orally, at a dosage level of from 5 up to about 100 mg per kilogram of host body weight.

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